Cytopathologic diagnosis of fine needle aspiration biopsies of thyroid nodules

Fine-needle aspiration(FNA) cytology is an important diagnostic tool in patients with thyroid lesions.Several systems have been proposed for the cyropathologic diagnosis of the thyroid nodules.However cases with indeterminate cytological findings still remain a matter of debate.In this review we analyze all literature regarding Thyroid Cytopathology Reporting systems trying to identify the most suitable methodology to use in clinical practice for the preoperative diagnosis of thyroid nodules.A review of the English literature was conducted,and data were analyzed and summarized and integrated from the authors' perspective.The main purpose of thyroid FNA is to identify patients with higher risk for malignancy,and to prevent unnecessary surgeries for benign conditions.The Bethesda System for Reporting Thyroid Cytopathology is the most widely used system for the diagnosis of thyroid FNA specimens.This system also contains guidelines for the diagnosis and treatment of indeterminate or suspicious for malignancy cases.In conclusion,patients who require repeated FNAs for indeterminate diagnoses will be resolved by repeat FNA in a percentage of 72%-80%.

TYMS/KRAS/BRAF molecular profiling predicts survival following adjuvant chemotherapy in colorectal cancer

BACKGROUND Patients with stage II-III colorectal cancer (CRC) treated with adjuvant chemotherapy, gain a 25% survival benefit. In the context of personalized medicine, there is a need to identify patients with CRC who may benefit from adjuvant chemotherapy. Molecular profiling could guide treatment decisions in these patients. Thymidylate synthase (TYMS) gene polymorphisms, KRAS and BRAF could be included in the molecular profile under consideration. AIM To investigate the association of TYMS gene polymorphisms, KRAS and BRAF mutations with survival of CRC patients treated with chemotherapy.METHODS A retrospective study studied formalin-fixed paraffin-embedded tissues (FFPEs) of consecutive patients treated with adjuvant chemotherapy during January/2005-January/2007. FFPEs were analysed with PCR for the detection of TYMS polymorphisms, mutated KRAS (mKRAS) and BRAF (mBRAF). Patients were classified into three groups (high, medium and low risk) according to 5’UTR TYMS polymorphisms Similarly, based on 3’UTR polymorphism ins/loss of heterozygosity (LOH) patients were allocated into two groups (high and low risk of relapse, respectively). Cox regression models examined the associated 5- year survival outcomes. RESULTS One hundred and thirty patients with early stage CRC (stage I-II: 55 patients;stage III 75 patients;colon: 70 patients;rectal: 60 patients) were treated with surgery and chemotherapy. The 5-year disease free survival and overall survival rate was 61.6% and 73.9% respectively. 5’UTR polymorphisms of intermediate TYMS polymorphisms (2RG/3RG, 2RG/LOH, 3RC/LOH) were associated with lower risk for relapse [hazard ratio (HR) 0.320, P = 0.02 and HR 0.343, P = 0.013 respectively] and death (HR 0.368, P = 0.031 and HR 0.394, P = 0.029 respectively). The 3’UTR polymorphism ins/LOH was independently associated with increased risk for disease recurrence (P = 0.001) and death (P = 0.005). mBRAF (3.8% of patients) was associated with increased risk of death (HR 4.500, P = 0.022) whereas mKRAS (39% of patients) not. CONCLUSION Prospective validating studies are required to confirm whether 2RG/3RG, 2RG/LOH, 3RC/LOH, absence of ins/LOH and wild type BRAF may indicate patients at lower risk of relapse following adjuvant chemotherapy.

Increased levels of circulating platelet-derived microparticles in psoriasis:Possible implications for the associated cardiovascular risk

AIM To evaluate platelet activation markers in psoriasis patients, compared to controls, and investigate their association with the inflammatory burden of psoriasis.METHODS Forty psoriatic patients without cardiovascular disease,and 12 healthy controls were subjected to measurement of baseline platelet CD62 P, CD63 and CD42 b expression, platelet-leukocyte complexes, i.e., platelet-monocyte complexes(PMC), platelet-neutrophil complexes(PNC) and platelet-lymphocyte complexes, and concentrations of platelet-derived microparticles(PMPs) using flow cytometry. Both larger-size(0.5-0.9 μm) and smallersize(< 0.5 μm) PMPs were determined. Serum interleukin(IL)-12 and IL-17 levels were also measured by enzyme-linked immunosorbent assay. The severity of psoriasis was evaluated by the Psoriasis Area Severity Index(PASI).RESULTS PMP concentrations were significantly higher in psoriasis patients than controls [mean±standard error of mean(SEM): 22±5/μL vs 11±6/μL; P=0.018), for both smaller-size(10±2/μL vs 4±2/μL; P=0.033) and larger-size(12±3/μL vs 6±4/μL; P=0.014) PMPs. Platelet CD62 P, CD63 and CD42 b expression and circulating PMC and PNC were similar between the two groups. Lower circulating PLC were observed in psoriasis patients compared to controls(mean±SEM: 16%±3% vs 23%±6%; P=0.047). Larger-size PMPs were related with IL-12 levels(P<0.001) and smaller-size PMPs with both IL-12 and IL-17 levels(P<0.001). Total PMPs also correlated with IL-12(P<0.001). CD63 expression was positively correlated with both IL-12 and IL-17(P<0.05). Increased PASI score was associated with increased levels of larger-size PMPs(r=0.45; P=0.011) and increased CD63 expression(r=0.47; P<0.01).CONCLUSION PMPs, known to be predictive of cardiovascular outcomes, are increased in psoriasis patients, and associated with high inflammatory disease burden. Enhanced platelet activation may be the missing link leading to cardiovascular events in psoriatic patients.

THE PHENOTYPE OF MAST CELL IN PRIMARY ADENOID LIVER TUMOR OF RAT AND ITS RELATION TO TUMOR CELL

Mast cells in adenoid liver tumors of 32 rats induced with nitrosomorpholine were observed ultrastructurally, and among them, some were studied immunocytochemically via immunogold techniques. Data indicating that mast cells which located in tumor tissues presented positive expression of rat mast cell protein (RMCP) Ⅰ, Indicating origination from the mucosa mast cells, while those in the connective tissues around tumors were largely stained negatively with either RMCP Ⅰor RMCP Ⅰ antisera, with the exception of only a few cells showing positive RMCP Ⅰ staining. Ultrastructural observation showed that mast cells in tumon contacted closely with the tumor cells. Membranes of the intracytoplasmic granules in these mast cells were fusing together. The content inside the granules were discharged and spread along the intercellular space between the tumor cells. There was not any lesion observed uitrasructrually in the tumor cells contacting with the mast cells. The significance of mucosa mast cells in adenoid liver tumors is briefly discussed.

Epithelial-Myoepithelial Carcinoma of Parotid Gland—A Surprising Clinical Entity

Objective: Objective of this study is to highlight the importance of considering clinical and radiological findings in parotid tumors before operating tumors with high suspicion of malignancy though reported as benign on histopathology. Material and Methods: 20 years old female patient underwent radical parotidectomy for epithelial-myoepithelial carcinoma of the left parotid gland which was diagnosed on FNAC. Contrast CT and contrast MRI also showed a tumor involving left parotid gland with destruction of ramus of mandible and loss of tissue planes medially & laterally. Facial nerve was infiltrated by tumor, thus sacrificed. Post operatively, patient had left facial palsy and was discharged after 7 days without any other sequelae. Postoperative histopathology showed pleomorphic adenoma with positive resection margins medially, superiorly & posteriorly. No further treatment was given in view of the benign nature of the tumor. Results: Patient was kept on monthly follow up and was disease free at one year follow up. Conclusion: Aggressive surgical treatment should be planned for malignant parotid tumors to give a better tumor free survival. Misdiagnosis can occur on FNAC as it can hit only one part of the tumor thus missing a malignant tumor or can interpret pleomorphic adenoma as epithelial-myoepithelial carcinoma.

Promoter Methylation, BRAF Mutation Analysis and Topoisomerase IIa Expression for the Detection of Endometrial Carcinoma in Liquid Based Cytology Samples

Cancer of the corpus uteri remains the most common gynecological related cancer in developed countries. Cytology, after the induction of liquid based cytology, has reemerged as a possible first line non-interventional diagnostic procedure with promising results. Apart from slide preparation for cytology diagnosis, LBC allows the application of elaborate molecular tests on the residual material. Samples from 74 symptomatic women were collected in ThinPrep?PreservCyt medium, from witch immunocytochemical and molecular tests were performed. Final diagnosis of 39 endometrioid carcinomas, 20 non-endometrioid carcinomas and 15 non-malignant was set after hysterectomy. Topoisomerase IIa expression was common (42%) in both types of cancer. Promoter methylation analysis revealed that hMLH1 is commonly methylated in cancers (52.7%), CDKN2A and MGMT less often (27.1%) and RARB rarely methylated (8.4%). BRAF activating mutation V600E was a rare event (8.4%) only found in low grade endometrioid carcinomas. Topoisomerase IIa expression correlated with BRAF mutations, hMLH1 and to lesser extent with CDKN2A methylation. Almost none of the biomarkers were positive in cytological negative or hyperplastic without atypia samples. Detection of methylation in any gene displayed sensitivity, specificity, PPV and NPV similar to cytology of cancer. However, inclusion of cytology diagnosis of hyperlasias with atypia increased sensitivity and NPV of cytology outperforming methylation of any gene. Further evaluation of the panel of promoter methylation, especially in cytology diagnoses of hyperplasia with or without atypia should be evaluated since initial results are promising. Even though methylation of MGMT and RARB are rare events, some patients could be benefit from specific chemotherapeutics that target either of them or the more frequently expressed topoisomerase IIa.

Analysis of Mortality Rates of the 5 Most Frequent Primary Cancer Sites in Brazil between 1979 and 2015 for Both Sexes

Introduction:Global estimates determined that there would be 14.1 million new cases of cancer and 8.2 million deaths from cancer.Objectives:To determine the adjusted mortality rates of the 5 most frequent primary sites of cancer and the mortality rates of the 5 most frequent primary in Brazil,between 1979 and 2015.Method:Epidemiological study between 1979 and 2015 based on data obtained from the Online Atlas of Mortality provided by the National Cancer Institute of Brazil(INCA)using information on cancer deaths obtained within the System of Information on Mortality(SIM)of Brazil Ministry of Health.Results:When analyzing the mortality rate of the 5 most frequent primary sites by type of cancer from 1979 to 2015 for both sexes,we show that in the year 2015 the cancers with the highest rates are:bronchi and lungs,breast,stomach,prostate and unknown primary location.A total of 527,980 deaths from bronchial and lung neoplasms were recorded,304,546 deaths from breast cancer were recorded,407,611 deaths from stomach cancer were recorded,268,061 deaths from prostate cancer were recorded,and 257,357 deaths due to neoplasia of unknown primary location were recorded.When we analyze the world standard rate and Brazil’s standard rate for mortality,we show that Brazil has higher values for the 5 most frequent primary sites by type of cancer.Conclusion:The epidemiological study of these values aims to determine what are the main descriptors and characteristics to determine preventive measures against the increase of this disease.

Diagnostic Value between 1984 and 2018 of Transrectal Biopsy Guided by Ultrasonography after Radical Prostatectomy

Objectives—to determine correlation between GSs(Gleason scores)on needle biopsy and RP(radical prostatectomy),evaluating diagnostic tests on biopsy and RP within the last years,between 1984 and 2018.Method—analysis of 100 patients,diagnosed with PCa(prostate cancer)needle biopsy using 18-gauge needle,who underwent RP with lymphadenectomy and for which preoperative and postoperative GSs were available.GS group analysis used three categorization schemes for differentiation:mild,moderate and poor for the whole group and we determined SE(sensitivity),SP(specificity),PVPR(positive predictive value),negative predictive value and accuracy.Results—we found that 42%of the patients had no changes between GS on biopsy and prostatectomy,while 20%were overgraded and 38%undergraded by needle biopsy.Graduation of+1 point in GS(32%)or-1 point(17%)was the most common.Most patients were classified as moderately differentiated by biopsies(78 and 35%in scheme 1 and 2 or 3,respectively),while 43%of patients received an intermediate differentiation classification.Biopsy accuracy varied from 44 to 76%for the analysis of all three schemes.Conclusion—there are differences in correlation between GS on biopsy and on surgical specimen,and Gleason’s graduation also depends on the experience of the pathologist.We have shown that sextant biopsies using 18-gauge and a same group of pathologists showed acceptable concordance values(42%)between the GS on biopsy and prostatectomy.

Integrating TYMS, KRAS and BRAF testing in patients with metastatic colorectal cancer

AIM To investigate the impact of thymidylate synthase(TYMS), KRAS and BRAF in the survival of metastatic colorectal cancer(m CRC) patients treated with chemotherapy. METHODS Clinical data were collected retrospectively from records of consecutive patients with m CRC treated with fluoropyrimidine-based chemotherapy from 1/2005 to 1/2007. Formalin-fixed paraffin-embedded tissues were retrieved for analysis. TYMS genotypes were identified with restriction fragment analysis PCR, while KRAS and BRAF mutation status was evaluated using real-time PCR assays. TYMS gene polymorphisms of each of the 3' untranslated region(UTR) and 5'UTR were classified into three groups according to the probability they have for high, medium and low TYMS expression(and similar levels of risk) based on evidence from previous studies. Univariate and multivariate survival analyses were performed.RESULTS The analysis recovered 89 patients with m CRC(46.1% de novo metastatic disease and 53.9% relapsed). Of these, 46 patients(51.7%) had colon cancer and 43(48.3%) rectal cancer as primary. All patients were treated with fluoropyrimidine-based chemotherapy(5FU or capecitabine) as single-agent or in combination with irinotecan or/and oxaliplatin or/and bevacizumab. With a median follow-up time of 14.8 mo(range 0-119.8), 85 patients(95.5%) experienced disease progression, and 63 deaths(70.8%) were recorded. The 3-year and 5-year OS rate was 25.4% and 7.7% while the 3-year progression-free survival rate was 7.1%. Multivariate analysis of TYMS polymorphisms, KRAS and BRAF with clinicopathological parameters indicated that TYMS 3'UTR polymorphisms are associated with risk for disease progression and death(P < 0.05 and P < 0.03 respectively). When compared to tumors without any del allele(genotypes ins/ins and ins/loss of heterozygosity(LOH) linked with high TYMS expression) tumors with del/del genotype(low expression group) and tumors with ins/del or del/LOH(intermediate expression group) have lower risk for disease progression(HR = 0.432, 95%CI: 0.198-0.946, P < 0.04 and HR = 0.513, 95%CI: 0.287-0.919, P < 0.03 respectively) and death(HR = 0.366, 95%CI: 0.162-0.827, P < 0.02 and HR = 0.559, 95%CI: 0.309-1.113, P < 0.06 respectively). Additionally,KRAS mutation was associated independently with the risk of disease progression(HR = 1.600, 95%CI: 1.011-2.531, P < 0.05). The addition of irinotecan in 1st line chemotherapy was associated independently with lower risk for disease progression and death(HR = 0.600, 95%CI: 0.372-0.969, P < 0.04 and HR = 0.352, 95%CI: 0.164-0.757, P < 0.01 respectively).CONCLUSION The TYMS genotypes ins/ins and ins/LOH associate with worst prognosis in m CRC patients under fluoropyrimidine-based chemotherapy. Large prospective studies are needed for validation of our findings.

Kawasaki disease, <i>Mycoplasma pneumoniae</i>infection and anaplastic large cell lymphoma: An uncommon association

Kawasaki disease (KD) is an acute febrile systemic vasculitis occurring predominantly in young children less than 5 years of age. Although imperfectly known, the aetiopathogenesis of KD would be secondary to immunological abnormalities that could constitute a favourable terrain for neoplasms. We report on a case in a 2-year-old girl who presented clinical manifestations compatible with Kawasaki disease complicated by coronary aneurysm. Aetiopathological investigations revealed M. pneumoniae infection as specific IgM were present in the serum (Elisa). The patient was initially treated by intravenous immunoglobulins (IVIG) and aspirin to anti-inflammatory dose. Following a few days of desquamation, resolution of the symptomatology occurred. Four weeks later she developed disseminated tumorous syndrome. Lymph node biopsy revealed massive infiltration by large cells lymphomatous proliferation. Histologic and immunophenotypic findings were characteristic of ALK-1+ anaplastic large cell lymphoma. Four weeks later, the patient died from a severe nosocomial infection complicated by septic shock. Our observation is the first cases describing the association between anaplastic large cell lymphoma, KD and M. pneumoniae. Immunologic disorder due to KD and M. pneumoniae infection may play probably a central role for malignancy.

Metastasis in an axillary lymph node in hepatocellular carcinoma:a case report

INTRODUCTIONAlthough hepatocellular carcinoma oftenmetastasizes to regional lymph nodes,spread tomore distant lymph nodes is rare.Involvementof axillary lymph nodes by metastases appears not tohave been documented.We report a patient withhepatocellular carcinoma (HCC) with a metastasisin a lymph node in the right axilla,and discusspossible routes by which such spread might occur.

Triglyceride and Glucose Index (TyG) is a Reliable Biomarker to Predict Non-Alcoholic Fatty Liver Disease

Non-alcoholic fatty liver disease (NAFLD) is a common liver disease worldwide. There is no specific biomarker for the diagnosis of NAFLD. Trigly-ceride and glucose index (TyG) may predict the subsequent occurrence of NAFLD in later life. This cross sectional study was aimed to evaluate the effectiveness of triglyceride and glucose index (TyG) as a possible biomarker of NAFLD. The study was conducted at the Department of Laboratory Medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from March 2019 to February 2020. A total of 124 subjects were taken as study population following selection criteria. Among them 62 were diagnosed patients of NAFLD and 62 were healthy subjects as control group. Fasting plasma glucose was measured by glucose oxidase method and serum triglyceride was measured by enzymatic-colorimetric method, while TyG index was calculated accordingly. The mean age was 39.5 ± 11.27 years in NAFLD patients and 37.10 ± 12.28 years in control subjects with male female ratio 1:1.7 and 1:1.8 respectively. Major portion of NAFLD patients (62.9%) were overweight (BMI ≥ 25). The mean fasting plasma glucose level was 5.73 ± 1.47 mmol/L in NAFLD patients and 5.27 ± 0.69 mmol/L in control group (p < 0.027). The mean serum triglyceride level was 237.19 ± 96.47 mg/dl in NAFLD patients and 117.32 ± 53.07 mg/dl in control group (p < 0.001). The triglyceride and glucose index (TyG) was 9.36 ± 0.47 in NAFLD group and 8.53 ± 0.42 in control group. TyG index was significantly higher in NAFLD patients in comparison to control group (p < 0.001). In ROC analysis, cut off value of TyG index was 8.85 with sensitivity 93.5% and specificity 79%. As a fast and effective method, TyG index can be used as a diagnostic tool to predict NAFLD.

Antibody-Cytokeratin Marker 34βE12 in Prostate Cancer Detection

Introduction:The histological diagnosis of prostate cancer is commonly based on morphological patterns.The presence of malignant tissue mixed with benign tissue,or the presence of carcinoma that mimics benignity may generate difficulty in the diagnostic elucidation.Therefore,the application of immunohistochemistry contributes its diagnostic value.Objectives:To evaluate the 34βE12 marker in the detection of adenocarcinoma(ADn),atypical small acinar proliferation(ASAp),regular prostatic tissue(RPT)and regular prostatic tissue alternated by atrophy spotlights(RPTa)in transrectal biopsy guided by ultrasonography of patients with suspected prostate cancer.Method:Analysis of 34 patients who underwent ultrasound-guided transrectal biopsy with subsequent analysis by H&E staining and 34βE12 labeling for elucidation of neoplasms or diseased tissues with doubtful diagnosis.Results:The marker 34βE12 showed negativity in 100%of the neoplasms ADn,positivity in 100%of the benign prostatic tissues(RPT and RPTa);the patients with ASAp presented positivity(20%)and negativity(80%).The chi-square test(χ2)showed that there is an association(χ2=29.55 and p<0.0001)between the groups,that is,the 34βE12 marker has a significant value(p<0.0001)in the elucidation of patients with prostatic neoplasia and benign prostatic tissues.Discussion and Conclusion:With the early screening of prostate cancer in the modern era,pathologists have become increasingly challenged to diagnose small outbreaks of cancer when only a few atypical glands are present in transrectal biopsy-guided ultrasonography.The 34βE12 marker becomes an important tool in elucidating diagnoses such as ADn and ASAp.

Mathematical modeling for Delta and Omicron variant of SARS-CoV-2 transmission dynamics in Greece

A compartmental,epidemiological,mathematical model was developed in order to analyze the transmission dynamics of Delta and Omicron variant,of SARS-CoV-2,in Greece.The model was parameterized twice during the 4th and 5th wave of the pandemic.The 4th wave refers to the period during which the Delta variant was dominant(approximately July to December of 2021)and the 5th wave to the period during which the Omicron variant was dominant(approximately January to May of 2022),in accordance with the official data from the National Public Health Organization(NPHO).Fitting methods were applied to evaluate important parameters in connection with the transmission of the variants,as well as the social behavior of population during these periods of interest.Mathematical models revealed higher numbers of contagiousness and cases of asymptomatic disease during the Omicron variant period,but a decreased rate of hospitalization compared to the Delta period.Also,parameters related to the behavior of the population in Greece were also assessed.More specifically,the use of protective masks and the abidance of social distancing measures.Simulations revealed that over 5,000 deaths could have been avoided,if mask usage and social distancing were 20%more efficient,during the short period of the Delta and Omicron outbreak.Furthermore,the spread of the variants was assessed using viral load data.The data were recorded from PCR tests at 417 Army Equity Fund Hospital(NIMTS),in Athens and the Ct values from 746 patients with COVID-19 were processed,to explain transmission phenomena and disease severity in patients.The period when the Delta variant prevailed in the country,the average Ct value was calculated as 25.19(range:12.32e39.29),whereas during the period when the Omicron variant prevailed,the average Ct value was calculated as 28(range:14.41e39.36).In conclusion,our experimental study showed that the higher viral load,which is related to the Delta variant,may interpret the severity of the disease.However,no correlation was confirmed regarding contagiousness phenomena.The results of the model,Ct analysis and official data from NPHO are consistent.